Immunofluorescence analysis of Syntrophin was done on 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Syntrophin (1351) Mouse Monoclonal Antibody (MA1745) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Amphibian, Dog, Chicken, Human, Mouse, Rat|
|Published species reactivity||Rat, Amphibian, Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Whole purified syntrophin from Torpedo californica electric organ postsynaptic membrane.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||0.5µg per 10^6 cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||0.1-1 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-745 detects alpha 1, beta 1 and beta 2 subunits of syntrophin. This antibody has been shown to react with human, mouse, rat, chicken, canine and amphibian samples.
MA1-745 has been successfully used in Western blot, immunoprecipitation and immunofluorescence procedures. By Western blot, this antibody detects an ~58 kDa protein representing syntrophin from rat and mouse cell extracts.
The MA1-745 immunogen is whole purified syntrophin from Torpedo californica electric organ postsynaptic membrane. Characterization studies of this antibody have demonstrated that clone 1351 is directed against an epitope within the PDZ domain of syntrophin. MA1-745, clone 1351, is seen as the "gold standard" for syntrophin assessment.
Syntrophins are structurally related proteins located at the neuromuscular synapse and play an important role in dystrophin associated protein (DAP) complex formation. To date there have been three distinct syntrophin isoforms discovered: alpha 1, beta 1, and beta 2. These proteins each contain a PDZ domain, a syntrophin-unique domain (SU), and three pleckstrin homology domains (PH1a,1b and 2). The PDZ domain has been shown to be responsible for binding syntrophin to nNOS and the DAP complex. This association results in localization of the nicotinic acetylcholine receptor to the cytoskeleton via the (S/T)XV C terminus.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Neuronal nitric oxide synthase localizes to utrophin expressing intercalated discs and stabilizes their structural integrity.
MA1-745 was used in immunohistochemistry - frozen section to study the localization of neuronal nitric oxide synthase to utrophin expressing intercalated discs and stabilization of their structural integrity
|Gonzalez JP,Crassous PA,Schneider JS,Beuve A,Fraidenraich D||Neuromuscular disorders : NMD (25:964)||2015|
Genetically induced dysfunctions of Kir2.1 channels: implications for short QT3 syndrome and autism-epilepsy phenotype.
MA1-745 was used in western blot to study a naturally occurring Kir2.1 mutation associated with short QT3 syndrome and autism-epilepsy
|Ambrosini E,Sicca F,Brignone MS,D'Adamo MC,Napolitano C,Servettini I,Moro F,Ruan Y,Guglielmi L,Pieroni S,Servillo G,Lanciotti A,Valvo G,Catacuzzeno L,Franciolini F,Molinari P,Marchese M,Grottesi A,Guerrini R,Santorelli FM,Priori S,Pessia M||Human molecular genetics (23:4875)||2014|
Human T-cell leukemia virus type 1 Tax protein interacts with and mislocalizes the PDZ domain protein MAGI-1.
MA1-745 was used in western blot to study the subcellular mislocalization of MAGI-1 by the Tax oncoprotein of HTLV-1 and the significance for the persistence of HTLV-1 infection
|Makokha GN,Takahashi M,Higuchi M,Saito S,Tanaka Y,Fujii M||Cancer science (104:313)||2013|
Absence of glial ¿-dystrobrevin causes abnormalities of the blood-brain barrier and progressive brain edema.
MA1-745 was used in western blot to investigate the role of glial alpha-dystrobrevin in the breakdown of the blood-brain barrier and progressive brain edema
|Lien CF,Mohanta SK,Frontczak-Baniewicz M,Swinny JD,Zablocka B,Górecki DC||The Journal of biological chemistry (287:41374)||2012|
Phosphorylation on threonine 11 of ß-dystrobrevin alters its interaction with kinesin heavy chain.
MA1-745 was used in western blot to study the role of beta-dystrobrevin threonine 11 phosphorylation in modulating its interaction with the kinesin heavy chain
|Fratini F,Macchia G,Torreri P,Matteucci A,Salzano AM,Crescenzi M,Macioce P,Petrucci TC,Ceccarini M||The FEBS journal (279:4131)||2012|
Differential requirement for utrophin in the induced pluripotent stem cell correction of muscle versus fat in muscular dystrophy mice.
MA1-745 was used in western blot to evaluate the stem cell therapy for muscular dystrophy mice
|Beck AJ,Vitale JM,Zhao Q,Schneider JS,Chang C,Altaf A,Michaels J,Bhaumik M,Grange R,Fraidenraich D||PloS one (6:null)||2011|
A high throughput screen identifies chemical modulators of the laminin-induced clustering of dystroglycan and aquaporin-4 in primary astrocytes.
MA1-745 was used in western blot to identify chemical modulators for laminin-induced clustering of dystroglycan and aquaporin-4
|Noël G,Stevenson S,Moukhles H||PloS one (6:null)||2011|
MLC1 trafficking and membrane expression in astrocytes: role of caveolin-1 and phosphorylation.
MA1-745 was used in western blot to study the role of caveolin-1 and phosphorylation in modulating the expression and trafficing of the MLC1 membrane protein
|Lanciotti A,Brignone MS,Camerini S,Serafini B,Macchia G,Raggi C,Molinari P,Crescenzi M,Musumeci M,Sargiacomo M,Aloisi F,Petrucci TC,Ambrosini E||Neurobiology of disease (37:581)||2010|
The 8th and 9th tandem spectrin-like repeats of utrophin cooperatively form a functional unit to interact with polarity-regulating kinase PAR-1b.
MA1-745 was used in western blot to investigate the mechanism of the interaction between utrophin and kinase PAR-1b
|Yamashita K,Suzuki A,Satoh Y,Ide M,Amano Y,Masuda-Hirata M,Hayashi YK,Hamada K,Ogata K,Ohno S||Biochemical and biophysical research communications (391:812)||2010|
Cardiac sodium channel Nav1.5 is regulated by a multiprotein complex composed of syntrophins and dystrophin.
MA1-745 was used in western blot to investigate the role of syntrophins and dystrophin on the proper expression and function of Na (v) 1.5
|Gavillet B,Rougier JS,Domenighetti AA,Behar R,Boixel C,Ruchat P,Lehr HA,Pedrazzini T,Abriel H||Circulation research (99:407)||2006|
Knockdown of MLC1 in primary astrocytes causes cell vacuolation: a MLC disease cell model.
MA1-745 was used in immunocytochemistry to develop a model of megalencephalic leukoencephalopathy with subcortical cysts (MLC) disease
|Duarri A,Lopez de Heredia M,Capdevila-Nortes X,Ridder MC,Montolio M,López-Hernández T,Boor I,Lien CF,Hagemann T,Messing A,Gorecki DC,Scheper GC,Martínez A,Nunes V,van der Knaap MS,Estévez R||Neurobiology of disease (43:228)||2011|
A role of tyrosine phosphatase in acetylcholine receptor cluster dispersal and formation.
MA1-745 was used in immunocytochemistry to investigate the regulation of AChR cluster dispersal and formation by PTPase
|Dai Z,Peng HB||The Journal of cell biology (141:1613)||1998|
Dystrophin-associated protein scaffolding in brain requires alpha-dystrobrevin.
MA1-745 was used in immunohistochemistry to investigate the role of alpha-dystrobrevin in brain protein scaffolding
|Bragg AD,Das SS,Froehner SC||Neuroreport (21:695)||2010|
Synaptic localization of neuroligin 2 in the rodent retina: comparative study with the dystroglycan-containing complex.
MA1-745 was used in immunohistochemistry to determine the localization of neuroligin 2 in rodent retinal synapses
|Lui L,Levinson JN,Noël G,Handrigan GR,Richman JM,El-Husseini A,Moukhles H||Journal of neuroscience research (88:837)||2010|
Syntrophins regulate alpha1D-adrenergic receptors through a PDZ domain-mediated interaction.
MA1-745 was used in immunoprecipitation to study the mechanism for syntrophin regulation of alpha1D-adrenergic receptors .
|Chen Z,Hague C,Hall RA,Minneman KP||The Journal of biological chemistry (281:12414)||2006|
Absence of alpha-syntrophin leads to structurally aberrant neuromuscular synapses deficient in utrophin.
MA1-745 was used in immunoprecipitation to investigate the impacts of the lack of alpha-syntrophin on neuromuscular synapses
|Adams ME,Kramarcy N,Krall SP,Rossi SG,Rotundo RL,Sealock R,Froehner SC||The Journal of cell biology (150:1385)||2000|